Modified extracellular vesicles that specifically target kidneys and methods of preparation and applications thereof

Abstract

The present invention discloses a method for the preparation and application of modified extracellular vesicles specifically targeted to the kidneys. These modified extracellular vesicles are derived from fibroblastic reticular cells in the mesentery tissue and express anti-inflammatory proteins after being modified with anchor peptides. By combining the extracellular vesicles secreted from fibroblastic reticular cells in the mesentery tissue with anchor peptides, the present invention ensures the effective targeting of the exosomes to the kidneys. Additionally, through genetic engineering of fibroblastic reticular cells, the exosomes secreted by these cells can carry the target molecule, anti-inflammatory protein CD5L, achieving precise targeted therapy for the kidneys. This innovative treatment strategy holds promising potential for the treatment of sepsis-associated acute kidney injury, bringing positive impacts to patients' health.

What is claimed is: 
     
         1 . A modified extracellular vesicle that specifically target kidneys, characterized in that the modified extracellular vesicle that specifically target the kidneys is obtained by co-incubating extracellular vesicles secreted by reticular fibroblasts from mesenteric tissue expressing CD5 antigen-like proteins with anchoring peptides. 
     
     
         2 . The modified extracellular vesicle that specifically target the kidneys according to  claim 1 , wherein the extracellular vesicles secreted by reticular fibroblasts from mesenteric tissue are obtained by isolating and culturing reticular fibroblasts derived from mesenteric tissue and extracting by centrifugation, and the positive membrane protein markers of the extracellular vesicles secreted by reticular fibroblasts from mesenteric tissue include HSP70, CD81, CD63, and CD9, while negative markers include GM130, Calexin, VDAC1, and TIMM23. 
     
     
         3 . The modified extracellular vesicle that specifically target the kidneys according to  claim 1 , wherein the anchoring peptides include KIM-1 targeting peptide. 
     
     
         4 . A preparation method for the modified extracellular vesicle that specifically target the kidneys according to  claim 1 , wherein the method includes:
 expressing an anti-inflammatory protein in fibroblastic reticular cells (FRC) to obtain anti-inflammatory protein-modified extracellular vesicles;   co-incubating the anti-inflammatory protein-modified extracellular vesicles with the anchoring peptides to obtain the modified extracellular vesicles that specifically target the kidneys, which is abbreviated as anti-inflammatory protein-LTH-FRC-Exos.   
     
     
         5 . The method according to  claim 4 , wherein a method of expressing the anti-inflammatory protein in FRC to obtain the anti-inflammatory protein-modified extracellular vesicles comprises:
 using CD5L gene shown in SEQ ID NO. 3 as a template, obtaining a gene fragment by PCR with primers shown in SEQ ID NO. 1-SEQ ID NO. 2, and inserting the gene fragment into BamHI and NotI restriction sites of PG-P1-VSVG plasmid by using enzyme cutting and ligation to obtain lentiviral plasmid pLenti-CD5L;   co-transfecting the lentiviral plasmid pLenti-CD5L with a packaging plasmid into 293T cells to obtain successfully packaged lentivirus, and then infecting reticular fibroblasts with the lentivirus to obtain an FRC cell line overexpressing CD5L through screening;   separating and centrifuging the FRC cell line overexpressing CD5L to obtain the anti-inflammatory protein-modified extracellular vesicles.   
     
     
         6 . A method of using the modified extracellular vesicles that specifically target kidneys according to  claim 1  in a preparation of drugs for the treatment and/or prevention of acute kidney injury in sepsis. 
     
     
         7 . A drug for the treatment and/or prevention of acute kidney injury in sepsis, characterized in that the drug comprises the modified extracellular vesicles that specifically target kidneys according to  claim 1 . 
     
     
         8 . The drug according to  claim 7 , wherein the drug further comprises a carrier or an excipient. 
     
     
         9 . A method for treating acute kidney injury in septic patients, characterized in that the method comprises administering the drug according to  claim 7 .